Lactobacillus Paragasseri LPG-9 Alleviates Nsaids-Induced Gastric Ulcers and Associated Coagulation Abnormalities and Anemia Via the IGF-1/PTEN/Akt/Foxo Signaling Pathway

Background: Peptic ulcers not only cause upper gastrointestinal bleeding but can also lead to severe anemia. The widespread use of NSAIDs is closely associated with the occurrence of these complications. In particular, the increased frequency of NSAID use among patients with chronic diseases such as aging and coronary heart disease significantly elevates the risk of gastric mucosal damage, bleeding, and anemia. Our research group previously isolated several probiotic strains from the gastric mucosal tissue of healthy individuals. Among these, a gastric-derived strain, Lactobacillus paragasseri LPG-9, was found to exhibit high colonization rates and good acid tolerance, showing potential in treating gastric diseases and preventing bleeding. However, the underlying mechanisms remain unclear. Therefore, this study aims to investigate the protective effects and potential mechanisms of LPG-9 on NSAID-induced gastric ulcers, coagulation abnormalities, and anemia in SD rats.

Methods: Thirty SD rats were randomly divided into three groups (control group, aspirin group, and LPG-9 intervention group). The rats were administered either physiological saline or LPG-9 (1x10^9 CFU) suspension via gavage. After seven consecutive days of gavage, the rats were given an aspirin suspension (500 mg/kg). Six hours post-treatment, the rats were sacrificed, and gastric tissues were collected. The samples were fixed in 4% paraformaldehyde for 24 hours and subjected to preprocessing (dehydration, embedding, sectioning, clearing, H&E staining, PAS staining, and sealing) and morphological observation. Western blot and ELISA assays were used to detect the expression levels of COX-1, COX-2, PGE2, SOD, GSH, CAT, and MDA, as well as changes in pro-inflammatory cytokines (IL-1β, TNF-α, IL-6) and anti-inflammatory cytokine (IL-10). Additionally, coagulation function indicators (such as coagulation factors and platelet count) and anemia-related indicators (such as hemoglobin and red blood cell count) were evaluated. The protective effects of LPG-9 pretreatment on gastric mucosal injury, bleeding, coagulation function, and anemia were analyzed. Statistical analysis was performed using SPSS 20.0 software, with P < 0.05 considered statistically significant.

Results: Macroscopic morphological observations revealed that the gastric mucosa of rats in the ulcer model group exhibited redness and swelling, visible ulcer surfaces, and a dark brown color, accompanied by numerous punctate and streaky hemorrhagic spots. Pretreatment with LPG-9 significantly inhibited aspirin-induced gastric injury in rats, markedly reducing the number of hemorrhagic spots. Rats in the aspirin-treated group showed significant gastric bleeding, coagulation dysfunction (reduced coagulation factor levels and platelet counts), and anemia (decreased hemoglobin and red blood cell counts). LPG-9 pretreatment notably improved these indices, alleviating gastric bleeding and anemia. H&E staining results indicated that aspirin caused severe gastric mucosal defects, local necrosis, and inflammatory cell infiltration. PAS staining results demonstrated that aspirin led to a significant thinning of the gastric mucosal mucus layer, which was markedly reversed by LPG-9 pretreatment. Compared to the aspirin group, LPG-9 pretreatment significantly increased the expression levels of COX-1, COX-2, PGE2, SOD, GSH, and CAT, and significantly decreased MDA levels, restoring them to normal levels. Additionally, elevated levels of pro-inflammatory cytokines (IL-1β, TNF-α, IL-6) and decreased levels of anti-inflammatory cytokine (IL-10) were significantly improved in the LPG-9 treatment group. Transcriptomic analysis suggested that LPG-9 ameliorates inflammatory response, antioxidative stress, reduces apoptosis, and maintains the integrity of the gastric mucosal barrier through the IGF-1/PTEN/Akt/FOXO signaling pathway. Non-targeted metabolomics results indicated that LPG-9 produces high levels of short-chain fatty acids and glutamine, which may further enhance its protective effects by promoting IGF-1 expression.

Conclusion: LPG-9 alleviates NSAIDs-induced gastric ulcers, coagulation abnormalities, and anemia by upregulating IGF-1 and activating the IGF-1/PTEN/Akt/FOXO signaling pathway, providing a novel therapeutic strategy for NSAIDs-related gastrointestinal injuries and complications.

No relevant conflicts of interest to declare.